Oxidative Stress-Mediated Overexpression of Uracil DNA Glycosylase in Leishmania donovani Confers Tolerance against Antileishmanial Drugs

Figure 8

Real-time PCR showing normalized transcription level of LdUNG. (a) WT cells treated with AmpB (0.125 ng/ml) and SAG (45 μM) for 8 hours. (b) Untreated WT, AmpB- and SAG-resistant clinical isolates. (c) AmpB-resistant and SAG-resistant clinical isolates treated with AmpB and SAG, respectively, for 8 hours. A housekeeping gene, α-Ldtubulin, was taken as internal control for normalising the variation in input. The results are expressed as mean ± SEM of three independent experiments. Data was analysed using one-way ANOVA, followed by Tukey’s multiple comparison test using GraphPad Prism 5.0. One-way ANOVA compares the relative fold change of LdUNG transcription levels (mean) of three unmatched groups of leishmania cells (control, AmpB-treated/AmpB-resistant/AmpB-resistant (AmpB-treated) and SAG-treated/SAG-resistant/SAG-resistant (SAG-treated)). The y-axis in all three bar graphs corresponds to relative fold change observed for transcriptional level of UNG in L. donovani cells under different treatment conditions. , , and . ns, nonsignificant. For the meaning of these statistical symbols, refer to ligand of Figure 1.

(a)

(b)

(c)